Liu J, Wu C, Bossie K, Bejaoui K, Hosler B A, Gingrich J C, Ben Hamida M, Hentati F, Schurr E, de Jong P J, Brown R H
Day Neuromuscular Research Laboratory, Massachusetts General Hospital East, Charlestown 02129, USA.
Genomics. 1998 Apr 1;49(1):23-9. doi: 10.1006/geno.1998.5204.
Miyoshi myopathy (MM) and limb-girdle muscular dystrophy subtype 2B (LGMD2B) map to the same region on chromosome 2p13. To facilitate the cloning of the defective gene causing these two diseases, we used a combination of chromosome walking and expressed sequence tag (EST) screening and identified 864 P1-derived artificial chromosomes (PACs) whose inserts map to the MM/LGMD2B candidate region and surrounding areas. Among them, 139 are from a chromosome 2-specific PAC library and 725 are from a total genomic PAC library. A 3-Mb contig spanning the candidate region for MM/LGMD2B was assembled. This contig contains 200 PACs, 10 known genetic markers, 5 new polymorphic markers, 57 sequence tagged sites (STSs) generated from PAC end fragments, and 4 random STSs. In addition, we mapped 24 ESTs to this contig and excluded 37 ESTs from the contig, thus eliminating them as candidate MM/LGMD2B genes. The high-resolution, sequence-ready PAC contig for the MM/LGMD2B region provides a backbone for the identification of the disease gene(s) and for clarification of the relationship between the two diseases.
三好肌病（MM）和2B型肢带型肌营养不良症（LGMD2B）定位于2号染色体p13区域的同一位置。为便于克隆导致这两种疾病的缺陷基因，我们结合了染色体步移法和表达序列标签（EST）筛选技术，鉴定出864个P1人工染色体（PAC），其插入片段定位于MM/LGMD2B候选区域及周边区域。其中，139个来自2号染色体特异性PAC文库，725个来自全基因组PAC文库。构建了一个跨越MM/LGMD2B候选区域的3兆碱基重叠群。该重叠群包含200个PAC、10个已知遗传标记、5个新的多态性标记、57个由PAC末端片段产生的序列标签位点（STS）和4个随机STS。此外，我们将24个EST定位到该重叠群，并将37个EST排除在重叠群之外，从而将它们排除在MM/LGMD2B候选基因之外。MM/LGMD2B区域的高分辨率、具备测序条件的PAC重叠群为鉴定致病基因和阐明这两种疾病之间的关系提供了一个框架。
